Bruno A. Benitez Laboratory

我们实验室的研究兴趣之一是集中在基因组数据中临床定义明确的患者队列的深层分子特征. 阿尔茨海默病(AD)和帕金森病(PD)是异质性疾病，根据症状的严重程度和优势，具有可识别的临床病理亚型. An accurate molecular profile could reduce clinical heterogeneity among AD and PD patients. 我们的实验室使用高通量和无假设的“组学”技术来解决这一重要的翻译方面, 包括下一代DNA测序, epigenomics, transcriptomics, proteomics, metabolomics, and lipidomics, to generate a highly detailed molecular atlas for AD and PD. We aim to create a framework for uncovering proteins, genes, pathways, 以及潜在的生物标记物，将提高我们对潜在疾病机制的理解, 预测病程, 改进临床试验的设计.

在黑质纹状体多巴胺能变性之外的多个大脑区域中，详细的细胞特异性转录组状态分子特征将为运动和非运动PD症状提供机制见解. 我们将整合神经病理学和snRNA-Seq数据，以确定PD患者不同脑区中不同的分子特征和/或共享的异常通路. 我们将整合PD和痴呆伴路易体(DLB)的snRNA-Seq数据和GWAS命中，以鉴定细胞类型特异性表达和数量性状位点，并确定基因富集因果通路中涉及的基因. 我们还将使用来自snRNA-seq的表达特征来优先考虑多基因全基因组位点下的致病基因. 我们期望在PD患者未被发现但受影响的大脑区域中发现细胞类型特异性差异表达基因(DEGs)，这将为理解PD的新细胞和分子机制提供基础.

多组学数据集的集成有助于揭示跨多个组学层的潜在机制. We will leverage multi-omics data to profile the heterogeneity of pathways contributing to PD. Our unique study design combining clinical and neuropathological data with genomics, epigenomics, transcriptomics, proteomics, 来自PD患者的代谢组学数据将识别与PD相关的分析物和具有区分特征的PD亚型. 我们将生成PD的详细分子景观，并破译神经生物学途径的遗传结构, linking these factors to PD and related dementias. In addition, 多组学数据可以识别新的和潜在的高价值治疗靶点，用于治疗和预防PD及相关痴呆.

我们已经成功地确定了AD与内溶酶体基因的常见和罕见变异之间的多种新的遗传关联(PLD3, RAB10, GRN12, TMEM106B, CPAMD8, MS4A4A). Recently, 我们进行了系统的, 综合评估每个内溶酶体基因编码变异对发生迟发性AD风险的贡献. 我们发现，几种溶酶体酶基因的杂合功能丧失(LoF)变异在AD患者中显著富集. 最大和最新的PD GWAS荟萃分析发现，几种溶酶体基因的常见变异与PD风险相关. 然而，目前还没有 in vitro or in vivo validation of most of those genetic associations. We performed intrastriatal inoculation of α-Syn PFFs in hemizygous and knockout (KO) GALC, ASAH1, and NAGLU 老鼠，以及野生型的幼崽. 这些研究旨在确定这些溶酶体酶在α-突触核蛋白(α-Syn)摄取中的作用, trafficking, aggregation, and clearance.

In 2011, we discovered that mutations in the DNAJC5 基因导致神经退行性疾病, the autosomal dominant form of neuronal ceroid lipofuscinosis (AD-ANCL). DNAJC5 encodes cysteine-string protein alpha (CSPα). 在随后的研究中, 我们在体外和体内证明了CSPα在内溶酶体中起生理作用. In recent years, 越来越多的体外证据表明，错误折叠相关蛋白分泌(MAPS)途径在α - syn等原聚集蛋白的细胞间传递中起作用, tau, and amyloid. α-Syn and tau secretion seem to be primarily mediated by MAPS. CSPα is a crucial regulator of MAPS by mediating the cargo entry to the late endosomes. However, α-Syn或淀粉样蛋白在体内的分泌和扩散是否依赖于dnajc5尚未确定. 我们使用CRISPR/ cas9校正的患者来源的诱导多能干细胞和原代细胞研究了DNAJC5变异在AD和PD发生风险中的作用 DNAJC5 on the secretion and spreading of α-Syn and amyloid.

1. Benitez BA, Belálcazar HM, Anastasía A, Mamah DT, Zorumski CF, Mascó DH, Herrera DG, de Erausquin GA. 在多巴胺能神经元中，sk3介导的电流的功能降低先于ampa受体介导的兴奋性毒性. Neuropharmacology. 2011年6月;60 (7 - 8):1176 - 86. PubMed PMID: 21044638; PubMed Central PMCID: PMC4301957.
2. Benitez BA， Alvarado D, Cai Y, Mayo K, Chakraverty S, Norton J, Morris JC, Sands MS, Goate A, Cruchaga C. Exome-sequencing confirms DNAJC5 mutations as cause of adult neuronal ceroid-lipofuscinosis. PLoS One. 2011;6(11):e26741. PubMed PMID: 22073189; PubMed Central PMCID: PMC3208569.
3. Benitez BA, Cooper B, Pastor P, Jin SC, Lorenzo E, Cervantes S, Cruchaga C. TREM2 is associated with the risk of Alzheimer's disease in Spanish population. Neurobiol Aging. 2013 Jun;34(6):1711.e15-7. PubMed PMID: 23391427; PubMed Central PMCID: PMC3596468.
4. Benitez BA, Cruchaga C. TREM2和神经退行性疾病. N Engl J Med. 2013年10月17日;369(16):1567-8. PubMed PMI: 24131187; PubMed Central PMCID: PMC4380008.
5. Benitez BA, Karch CM, Cai Y, Jin SC, Cooper B, Carrell D, Bertelsen S, Chibnik L, Schneider JA, Bennett DA, Fagan AM, Holtzman D, Morris JC, Goate AM, Cruchaga C. The PSEN1, p.E318G variant increases the risk of Alzheimer's disease in APOE-ε4 carriers. PLoS Genet. 2013;9(8):e1003685. PubMed PMID: 23990795; PubMed Central PMCID: PMC3750021.
6. Benitez BA, Jin SC, Guerreiro R, Graham R, Lord J, Harold D, Sims R, Lambert JC, Gibbs JR, Bras J, Sassi C, Harari O, Bertelsen S, Lupton MK, Powell J, Bellenguez C, Brown K, Medway C, Haddick PC, van der Brug MP, Bhangale T, Ortmann W, Behrens T, Mayeux R, Pericak-Vance MA, Farrer LA, Schellenberg GD, Haines JL, Turton J, Braae A, Barber I, Fagan AM, Holtzman DM, Morris JC, Williams J, Kauwe JS, Amouyel P, Morgan K, Singleton A, Hardy J, Goate AM, Cruchaga C. Missense variant in TREML2 protects against Alzheimer's disease. Neurobiol Aging. 2014 Jun;35(6):1510.e19-26. PubMed PMID: 24439484; PubMed Central PMCID: PMC3961557.
7. Benitez BA， Davis AA, Jin SC, Ibanez L, Ortega-Cubero S, Pastor P, Choi J, Cooper B, Perlmutter JS, Cruchaga C. 帕金森病全基因组关联研究中5个孟德尔基因和顶级基因的重测序分析. Mol Neurodegener. 2016 Apr 19;11:29. PubMed PMID: 27094865; PubMed Central PMCID: PMC4837564. *通讯作者
8. Benitez BA, Sands MS. 来自CSPα突变携带者的原代成纤维细胞重现了成人发病的神经性神经样脂褐质病的特征. Sci Rep. 2017年7月24日;7(1):6332. PubMed PMID: 28740222; PubMed Central PMCID: PMC5524943. *通讯作者
9. Ibanez L, Dube U, Davis AA, Fernandez MV, Budde J, Cooper B, Diez-Fairen M, Ortega-Cubero S, Pastor P, Perlmutter JS, Cruchaga C, Benitez BA*. Pleiotropic Effects of Variants in Dementia Genes in Parkinson Disease. Front Neurosci. 2018 Apr 10;12:230. eCollection 2018. PMID: 29692703 PubMed Central PMCID: PMC5902712. *通讯作者
10. Deming Y, Filipello F, Cignarella F, Cantoni C, Hsu S, Mikesell R, Li Z, Del-Aguila JL, Dube U, Farias FG, Bradley J, Budde J, Ibanez L, Fernandez MV, Blennow K, Zetterberg H, Heslegrave A, Johansson PM, Svensson J, Nellgård B, Lleo A, Alcolea D, Clarimon J, Rami L, Molinuevo JL, Suárez-Calvet M, Morenas-Rodríguez E, Kleinberger G, Ewers M, Harari O, Haass C, Brett TJ, Benitez BA^， Karch CM^， Piccio L^， Cruchaga C^. The MS4A gene cluster is a key modulator of soluble TREM2 and Alzheimer's disease risk. Sci Transl Med. 2019 Aug 14;11(505)PubMed PMID: 31413141; PubMed Central PMCID: PMC6697053. ^Equal contribution
11. Li Y, Xu Y, Benitez BA, Nagree MS, Dearborn JT, Jiang X, Guzman MA, Woloszynek JC, Giaramita A, Yip BK, Elsbernd J, Babcock MC, Lo M, Fowler SC, Wozniak DF, Vogler CA, Medin JA, Crawford BE, Sands MS. 克拉布病中酸性神经酰胺酶的基因消融证实了精神病假说，并确定了新的治疗靶点. 美国国家科学基金委. 2019 Oct . 1;116(40):20097-20103. PubMed PMID: 31527255; PubMed Central PMCID: PMC6778236.
12. Mikulka CR, Dearborn JT， Benitez BA, Strickland A, Liu L, Milbrandt J, Sands MS. Cell-autonomous expression of the acid hydrolase galactocerebrosidase. 美国国家科学基金委. 2020年4月21日;17(16):9032-9041. PubMed PMID: 32253319; PubMed Central PMCID: PMC7183170
13. Ibanez L, Bahena JA, Yang C, Dube U, Farias FHG, Budde JP, Bergmann K, Brenner-Webster C, Morris JC, Perrin RJ, Cairns NJ, O'Donnell J, Álvarez I, Diez-Fairen M, Aguilar M, Miller R, Davis AA, Pastor P, Kotzbauer P, Campbell MC, Perlmutter JS，王志强，王志强，王志强, Benitez BA*. 功能基因组分析揭示apoe介导的帕金森病脑和脑脊液β -淀粉样蛋白水平调节. 神经病理学杂志. 2020 Nov 19;8(1):196. PubMed PMID: 33213513; PubMed Central PMCID: PMC7678051. *通讯作者
14. Davis AA, Inman CE, Wargel ZM, Dube U, Freeberg BM, Galluppi A, Haines JN, Dhavale DD, Miller R, Choudhury FA, Sullivan PM, Cruchaga C, Perlmutter JS, Ulrich JD, Benitez BA， Kotzbauer PT, Holtzman DM. APOE genotype regulates pathology and disease progression in synucleinopathy. Sci Transl Med. 2020 Feb 5;12(529)PubMed PMID: 32024799; PubMed Central PMCID: PMC7289511
15. Yang C, Farias FHG, Ibanez L, Suhy A, Sadler B, Fernandez MV, Wang F, Bradley JL, Eiffert B, Bahena JA, Budde JP, Li Z, Dube U, Sung YJ, Mihindukulasuriya KA, Morris JC, Fagan AM, Perrin RJ, Benitez BA，王志强，王志强，王志强. 大脑蛋白质组的基因组图谱, CSF and plasma prioritizes proteins implicated in neurological disorders. Nat Neurosci. 2021年9月,24 (9):1302 - 1312. PMID: 34239129.
16. Brase L, You SF, del Aguila J, Dai Y, Novotny BC, Soriano-Tarraga C, Dykstra T, Fernandez MV, Budde JP, Bergmann K, Morris JC, Bateman RJ, Perrin RJ, McDade E, Xiong C, Goate A, Farlow M, Chhatwal JP, Schofield P, Chui H, D. I. A. N. (DIAN)， Sutherland GT, Kipnis J, Karch CM^， Benitez BA^, Cruchaga C^, Harari O, A landscape of the genetic and cellular heterogeneity in Alzheimer disease, medRxiv , 2021.11.30.21267072 (2021). ^资深作者等额贡献
17. Novotny, BC, Fernandez, MV, Bahena, J, Budde, JP, Bergmann, K, Eteleeb, A, Bradley, J, Webster, C, Ebl, C, Norton, J, Gentsch, J, Dube, U, Wang, F, Morris, JC, Bateman, RJ, Perrin, RJ, McDade, E, Xiong, C, Chhatwal, J, Goate A, Farlow M, Schofield P, Chui H, Karch CM, Benitez BA^, Cruchaga C^, Harari O, the DIAN(DIAN) Group. 常染色体显性和晚发性阿尔茨海默病大脑的代谢组学和脂质组学特征. MedRxiv, 2021.11.04.21265941. ^资深作者等额贡献

我们实验室项目的资金来自美国国立卫生研究院. We are also sincerely grateful to the patients, family members, 以及贡献时间的护理人员, brains, 以及用于生物医学研究的生物标本.

Our lab is located in the Longwood Medical Area in the Center for Life Sciences 6th Floor, room #630, surrounded by collaborators from the Departments of Neurology and Medicine at 金宝搏手机登录.